Process of isolating the active principle of the suprarenal glands.



7 NEED STATES PATENT OFFICE- Patented June 2, 1903.

J OKICHI TAKAMINE, OF YORK, N. Y.

PROCESS OF ISOLATING THE ACTIVE PRlNClPLE OF THE SUPRARENAL GLANDS.

SPECIFICATION forming part of Letters Patent No. 730,196, dated une 2,1963.

Original application filed November 5, 1900, Serial No. 35,546. Dividedand this application filed November 26, 1900. Serial No. 37,729. (Nospecimens.)

To all whom, it may concern:

Be it known that I, J OKICHI TAKAMINE, a subject of the Emperor ofJapan, residing in the city of New York,county and State of New York,have invented certain new and useful Improvements in Processes ofObtaining a New and Useful Product of the Suprarenal Glands; and I dohereby declare the following to be a full, clear, and exact descriptionof the invention, such as will enable others skilled in the art to whichit appertains to make and use the same.

This invention relates to improvements in processes of obtaining a newand useful product possessing in a stable and concentrated 7 weight ofWater or other solvent and steeped at i a temperature of about 60 to 75centigradefor the period of about five to ten hours in a suitablevessel to obtain a fluid extract of the glands, preferably avoidingcontact with atmospheric air, which can be to a great extentaccomplished by a film of fat floating on top of the liquid, or may bedone by passing a slow current of hydrogen or carbon dioxid into the toppart of the vessel. The object of this is to prevent the oxidation ofthe extract. At the latter part of the steeping the temperature of themixture may be raised from 85 to 100 centigrade. The mass is nowstrained, and the residueis pressed to squeeze 0 at as much liquid aspossible. The residue thus pressed out is steeped again with the leastamount of water to cover the mass for several hours at the sametemperature as above. The two extracts thus obtained are mixed, and themixture is cooled rapidly and the solidified fat removed. The liquid isnow evaporated at a low temperature, preferably in a vacuum-pan,admitting, if necessary, a small quantity of hydrogen or carbon dioxidto replace air and to prevent oxidation. The liquidis evaporated untilit becomes one-fifth to one-seventh of the original volume. To thisconcentrated solution two to three times its own volume of alcohol isadded, so that the mixture will contain about sixty per cent. of alcoholby volume. This addition of alcohol will precipitate nonactive bodies,such as albuminoids, phosphates, and other mineral salts. filtered andthe filtrate distilled under vacuum to separate the alcohol. Thefiltrate is thus concentrated to such a degree that its specific gravitybecomes 1.95 to 1.15, and then caustic ammonia (ammonium hydrate) isadded until the solution acquires a strong smell of ammonia. Thesolution is then allowed to remain in a cool place for twelve totwentyfour hours,-When formation of the crystals will be observed. Thecrystalline substance so produced is filtered and washed with water andalcohol and dried and constitutes an active alkaloid principle of thesuprarenal gland and is the product sought.

In order to further purify the product, it may be dissolved in the leastpossible amount of Weak acid, such as acetic acid or hydrochloric orother acids. The solution is then filtered, if necessary, and causticammonia or alkaline carbonates are added, so as to make the solutionstrongly alkaline. The product Will then crystallize out in much purerform than before, being practically free from inorganic matter. Thisprocess of recrystallization may be repeated until the substance becomesquite pure, or the crude substance may be dissolved in the leastpossible amount of weak caustic alkali, filtered, if

necessary, and carbon dioxid then passed into i the solution, therebyforming a crystalline precipitate of the substance, thisrecrystallization being repeated, if desired.

During the above-described manipulations It is then oxidation of thesubstance by the absorption of oxygen from the air must be avoided asfar as possible, due precautions being taken for that purpose.

In refining the material by the use of either acid or alkali I prefer,after dissolving it in such acid or alkali, to add strong alcohol in theproportion of from two to ten times its own volume to precipitate themineral impurities contained therein. The liquid is then filtered, andto the clear filtrate about one-half its own volume of ether ispreferably added, this precipitating dark-brown foreign matters, chieflyconsisting of coloringmatters, which are then removed by filtration. Thefiltrate is then neutralized with alkaline carbonates or ammonia orsuitable alkaline substances, if acid has been employed, or with an acidreagent, if the solution has been effected with alkali, and thesubstance is thereby precipitated.

In either method of purification the crystalline precipitate isfiltered, washed with water and alcohol, and dried.

The method of purification in which caustic alkali and a neutralizingagent is employed is not specifically claimed herein, as it forms thesubject-matter of another divisional application, Serial No. 30,730.

Having thus described my invention, the following is what I claim as newtherein and desire to secure by Letters Patent:

1. The process of obtaining a product of the suprarenal glands, whichconsists in making a fluid extract of said glands, precipitating fromsaid extract the inert albuminoid and mineral matters, removing saidprecipitate, treating the residue with ammonia to precipitate the saidproduct.

2. The process of obtaining a product of the suprarenal glands whichconsists in making an aqueous extract of said glands, precipitating fromsaid extract inert albuminoids and mineral matters, removing theprecipitate, adding ammonia to the residue to precipitate the saidproduct therefrom.

3. The process of obtaining a product of the suprarenal glands, whichconsists in concentrating the aqueous extract of the fresh gland, and inadding alcohol to precipitate inert albuminoids and mineralmatters,evaporating the alcohol and water so as to further concentratethe liquid, then adding ammonia in sulficient quantity to precipitatethe said product in alkaloid form, washing the precipitate with suitableliquid, and drying the same, all substantially as set forth.

4-. The process of obtaining a product of the suprarenal glands, whichconsists in steeping in water the comminuted glands at a suitabletemperature for a suitable number of hours; separating the solublematter by filtration and pressing; evaporating the filtrate to suchconsistency that the inert albuminoid and mineral salts will precipitateout by addition of a suitable amount of alcohol, adding a suitableamount of alcohol to precipitate the maximum amount of inert albuminoidand mineral matters, evaporating oil? the alcohol, and furtherevaporating the liquid to a suitable strength; adding ammonia insufficient quantity to precipitate the said product in crystalline form,separating the crystalline precipitate, washing with water and alcohol,and drying the product, all substantially as described.

5. The process of obtaining a product of the suprarenal glands, whichconsists in steeping in water the comminuted glands at a suitabletemperature for a suitable number of hours, separating the solublematter by filtration and pressing, evaporating the filtrate to suchconsistency that the inert albuminoid and mineral salts will precipitateout by addition of a suitable amount of alcohol, adding a suitableamount of alcohol to precipitate the maximum amount of inert albuminoidand mineral matters, distilling of? alcohol, and further evaporating theliquid to a suitable strength, adding ammonia in sufficient quantity toprecipitate the said product of the glands ina crystalline form, Washingthe precipitate with water and alcohol and drying the product, allsubstantially as described.

6. The process of refining and preparing the product of the suprarenalglands in purer form, which consists in dissolving said prodnot in asuitable acid solvent, adding to said solution strong alcohol toprecipitate impurities therefrom, separating the said precipitatedimpurities, treating the residue with an alkaline reagent to neutralizethe sol vent and precipitate the product, washing said preciph tatedactive principle with alcohol and water and then drying the same.

7. The process of refining and preparing the product of the suprarenalglands in purer form, which consists in dissolving said product indilute acetic acid, adding to said solution strong alcohol toprecipitate impurities therefrom, separating the said precipitatedimpurities, treating the residue with an alkaline reagent to neutralizethe solvent and precipitate the active principle, washing saidprecipitated product with alcohol and water and then drying the same.

8. The process of refining and preparing the product of the suprarenalglands in purer form, which consists in dissolving the said product in anon-neutral solvent, adding alcohol to said solution to precipitatemineral impurities, separating said precipitated im purities, addingether to the residue to pre cipitate coloring-matters, separating saidprecipitated coloring-matters, and adding to the residue a neutralizingagent for a solvent to precipitate the said product.

9. The'process of refining and preparing oring-matters, and adding tothe residue a the product of the suprarenal-glands in purerneutralizingagent for a solvent to precipitate (0 form, which consists in dissolvingsaid prodthe said product. i

not in an acid solvent, adding alcohol to said 5 solution to precipitatemineral impurities, JOKICHI TAKAMINE' separating said precipitatedimpurities, add- Witnesses: ing ether to the residue to precipitatecolor- J. GREEN, ing-matters, separating said precipitated coll A.-P.KNIGHT.

